[No public or meaningful name is available]

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames assay:

The test substance did not induce gene mutation in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA100 and TA98 in the presence and absence of S9 metabolic activation system by plate-incorporation assay and hence is not likely to classify as a gene mutant in vitro.

 

In vitro mammalian chromosome aberration study:

The test chemical did not induce chromosome aberrations in the mammalian cell line in the presence and absence of S9 metabolic activation system and hence it is not mutagenic in the chromosome aberration study performed.

 

In vitro gene mutation study in mammalian cells:

Test chemical did not induce mutation in mammalian cell line in the presence and absence of metabolic activation and hence it is not likely to classify as a gene mutant in vitro.

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data is from secondary source

Qualifier:

according to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

GLP compliance:

not specified

Type of assay:

in vitro mammalian cell gene mutation test using the Hprt and xprt genes

Specific details on test material used for the study:

Not specified

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Details on mammalian cell type (if applicable):

Not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 metabolic activation

Test concentrations with justification for top dose:

0.1 to 4.0 mg/ml without S9; 2.0 to 5.0 mg/ml with S9 activation

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

N-dimethylnitrosamine

methylmethanesulfonate

Details on test system and experimental conditions:

Dose Selection - Appropriate concentrations for mutagenicity testing were determined by preliminary measurements of cytotoxicity to CHO cells of a range of concentrations tested both in the presence and absence of a rat-liver S9 metabolic activation system. Selection of a suitable range of concentrations for testing was based upon an estimate of the doses which would not produce excessive cytotoxicity to the treated cells. Dimethylsulfoxide (DMSO) was used as the solvent for dilutions. All dilutions were prepared immediately prior to testing.
Test Procedure - Duplicate cultures of CHO cells were exposed for 5 hours to a minimum of five concentrations of Organofunctional Silane A-1120 in test both with and without the addition of a rat-liver S9 metabolic activation system. Various dose levels of Organofunctional Silane A-1120 for testing were attained by direct addition of various aliquots of the diluted test agent into the cell culture medium. The surviving fraction was determined at 18 to 24 hours after the removal of the test chemical using 4 plates/culture and 100 cells/plate. The mutant fraction was determined after a 9 to 12 day sub-culturing period to allow "expression" of the mutant phenotype. The mutant fraction was assessed in selective medium with 2 x 10E5 cells/plate in 5 plates/dosed culture (i.e. 1 x 10E6 total cells/dosed culture). The plating efficiency of these cells was assessed in non-selective medium using 4 plates/dosed culture with 100 cells/plate. The mutagenicity/survival/plating efficiency data from at least the top five concentrations which allowed sufficient cell survival for assessment of survival and quantification of mutants were recorded. The percentage of cells surviving the treatment, the numbers of mutant colonies, the percentage of clonable cells and the calculated number of mutants/10E6 clonable cells were presented. Positive (dimethylnitrosamine with S9 activation and ethylmethanesulfonate without S9 activation) and vehicle control (cell culture medium and DMSO) materials were tested concurrently to assure both the sensitivity of the test system.

Statistics:

The data were analyzed in comparison to concurrent control values after transformation of the mutation frequencies (MF) and SCE values according to the conversion method of Box and Cox (1964). This procedure for CHO data follows procedures described by Snee and Irr: (MF + 1)^0.15 (Snee, R.D. and J.D. Irr, Mutation Research, 85 (1981), 77-93). Data for positive and negative controls were compared to historical ranges but were not analyzed statistically.

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

6 mg/ml and higher in tests with and without S9 activation

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: No mutagenic potential was observed

Conclusions:

The test substance did not induce any statistical significant increase in the number of cells in the presence or absence of the S9 metabolic activation. Hence, the test substance can be considered to be non-mutagenic.

Executive summary:

The test chemical was assayed for gene mutations in the Chinese Hamster Ovary (CHO) cells both in the absence and presence of S9 metabolic activation. Positive (dimethylnitrosamine with S9 activation and ethylmethanesulfonate without S9 activation) and vehicle control (cell culture medium and DMSO) materials were tested concurrently to assure both the sensitivity of the test system. The study was performed according to OECD 476 Guidelines. The test concentrations used were- 0.1 to 4.0 mg/ml without S9 metabolic activation and 2.0 to 5.0 mg/ml with S9 activation. did not product any statistically significant increases in the incidence of mutations of CHO cells within a range of cytotoxic-to-noncytotoxic concentrations between 2.5 to 4.0 mg/ml in tests without an S9 metabolic activation system. With S9 activation, one intermediate dose of 2.5 mg/ml produced a mutant incidence in one of the two dosed cultures which was statistically greater than the concurrent controls. No dose-related trend in mutant values was observed in the test with or without S9 activation. The biological significance of the single increase was evaluated by determining reproducibility in an independent repeat test over a narrower range of concentrations with S9 activation. No significant or dose-related increases were observed in the repeat test. The test substance did not induce any statistical significant increase in the number of cells in the presence or absence of the S9 metabolic activation. Hence, the test substance can be considered to be non-mutagenic.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data is from peer reviewed publication

Qualifier:

according to guideline

Guideline:

other: Refer below principle

Principles of method if other than guideline:

In vitro mammalian chromosomal aberration test was performed to determine the mutagenic nature of the test chemical

GLP compliance:

not specified

Type of assay:

other: In vitro mammalian chromosomal aberration test

Target gene:

No data

Species / strain / cell type:

mammalian cell line, other: A pseudo-diploid Chinese hamster cell line (Don)

Details on mammalian cell type (if applicable):

- Type and identity of media: Eagle's minimum essential medium supplemented with 10% fetal calf serum (pH 7.2) adjusted by HEPES6 buffer
- Properly maintained: Yes
- Periodically checked for Mycoplasma contamination: Yes
- Periodically checked for karyotype stability: Yes, CHL cells had modal chromosome numbers of 21 and 25
- Periodically "cleansed" against high spontaneous background: No data

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

No data

Metabolic activation:

with and without

Metabolic activation system:

rat liver S-9 induced with Aroclor 1254

Test concentrations with justification for top dose:

0, 0.000001, 0.00001, 0.0001 or 0.001 M

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium
Cells at the start of seeding: 1.0-1.2 X 106 cells per TD-40 culture bottle

DURATION
- Preincubation period: No data
- Exposure duration: 26 hrs
- Expression time (cells in growth medium): 26 hrs
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): aqueous solution of 33258 Hoechst and Giemsa

NUMBER OF REPLICATIONS: Duplicate

NUMBER OF CELLS EVALUATED: 100 metaphase plates for each dose

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: Yes, mitotic index

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: The chromosome slide was stained in aqueous solution of 33258 Hoechst for 10 minutes, rinsed briefly in tap water, and mounted in phosphate buffer (pH 7.0) with a cover slip. The slide was exposed to an electric light (60 W, at 12-cm distance) for 1 hour. The cover slip was removed by tap water, and the slide was incubated in 1 M NaH2P04 (pH 8.0, 83-85° C) for 10 minutes, rinsed, and stained in 2.5% Giemsa (in phosphate buffer, 0.07 M, pH 7.0) for 5 minutes. Conventional Giemsa-stained slides were also prepared for scanning of chromosome aberrations.

Rationale for test conditions:

No data

Evaluation criteria:

The frequency of aberrations, excluding gaps, was indicated by the number of breaks per cell. A ring, a dicentric, and a chromatid exchange were each scored as two breaks, a tricentric as four breaks, and an acentric fragment or an isochromatid break as one break.

Statistics:

No data

Species / strain:

mammalian cell line, other: CHL

Metabolic activation:

not specified

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

at 0.00001 and 0.0001 M

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

True negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

No data

Remarks on result:

other: No mutagenic potential

Table: Frequencies of chromosome aberrations in Don cells after treatment with BUdR and solvent

Treatment	No. of cultures	No. of cultures observed for	Breaks/cell
			Mean±SE	Range
BUdR + DMSO	6	600	0.0666±0.0010	0.02- 0.11

 

TABLE 2. Chromosome aberrations in Don cells exposed to chemical

Chemical	Dose (M)	MI	Breaks/cell
Test chemical	0.000001	-	0.02
	0.00001	-	0.09
	0.0001	+	0.03
	0.001	++	-

Conclusions:

The test chemical did not induce chromosome aberration in pseudo-diploid Chinese hamster cell line (Don) and hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

In vitro mammalian chromosomal aberration test was performed to determine the mutagenic nature of the test chemical. The test chemical was mixed with DMSO and used at dose level of 0, 0.000001, 0.00001, 0.000, or 0.001 M using pseudo-diploid Chinese hamster cell line (Don). Three hours after 1.0-1.2 X 106 cells per TD-40 culture bottle were seeded, BUdR (1µg/ml) and test chemical was added to the cultures under an ordinary yellow darkroom safety lamp. Concurrent solvent control was also included in the study. All cultures were kept in complete darkness at 37° C for 26 hours (this covered two rounds of cell cycle), and 0.25µg colchicine/ml was added for the final 2 hours. The cells were collected by scraping them with a rubber policeman and prepared air-dried slides following hypotonic treatment and fixation in ice-cold methanol: acetic acid (3: 1). The chromosome slide was stained in aqueous solution of 33258 Hoechst for 10 minutes, rinsed briefly in tap water, and mounted in phosphate buffer (pH 7.0) with a cover slip. The slide was exposed to an electric light (60 W, at 12-cm distance) for 1 hour. The cover slip was removed by tap water, and the slide was incubated in 1 M NaH2P04 (pH 8.0, 83-85° C) for 10 minutes, rinsed, and stained in 2.5% Giemsa (in phosphate buffer, 0.07 M, pH 7.0) for 5 minutes. Conventional Giemsa-stained slides were also prepared for scanning of chromosome aberrations. The frequency of aberrations, excluding gaps, was indicated by the number of breaks per cell. A ring, a dicentric, and a chromatid exchange were each scored as two breaks, a tricentric as four breaks, and an acentric fragment or an isochromatid break as one break. The test chemical did not induce chromosome aberration in pseudo-diploid Chinese hamster cell line (Don) and hence it is not likely to classify as a gene mutant in vitro.

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Study period:

Not specified

Reliability:

4 (not assignable)

Justification for type of information:

Data was taken from secondary source

Principles of method if other than guideline:

Bacterial reverse mutation assay was carried out to determined the genetic toxicity of the given test chemical.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Not specified

Target gene:

Histidine operon (S. typhimurium strains); tryptophan operon (E. coli).

Species / strain / cell type:

other: Salmonella typhimurium strains TA 98, TA 100, TA 1535 and TA 1537

Details on mammalian cell type (if applicable):

Not specified

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

Not specified

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital and ß-Naphthoflavone induced rat liver S9

Test concentrations with justification for top dose:

0, 50, 160, 500, 1600 and 5000 µg/plate

Vehicle / solvent:

Vehicle(s)/solvent(s) used: DMSO

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other:

Remarks:

All active strains

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA 1537 (without activation)

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

TA 100, TA 1535 (without activation)

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA 98 (without activation)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION

- Preincubation period: 30 minutes

- Exposure duration: 72 hours

- Expression time (cells in growth medium): 72 hours

NUMBER OF REPLICATIONS: 3 plates per test concentration

DETERMINATION OF CYTOTOXICITY

- Method: relative total growth; background lawn assessment

Rationale for test conditions:

Not specified

Evaluation criteria:

For a test compound to be considered positive, it must (in two independent experiments) cause at least a doubling in the mean revertants per plate of at least one tester strain. This increase must be accompanied by a dose response towards increasing concentrations of the test article. Single increases in revertant frequencies, which are not dose-related and not reproducible in two independent tests are considered non-relevant. If however these increases do occur in both tests, this will be taken as an indication of mutagenic effect.

Statistics:

Mean number of revertants per plate and the standard deviation around the mean were calculated.

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: >5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: >5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: >5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: >5000 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: Numbers of spontaneous revertants were within acceptable range

Remarks on result:

other: No mutagenic potential

	TA98			TA100			TA1535
Conc. (µg/plate)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)
0*	14	30	No	139	151	No	8	9	No
50	16	37	No	147	144	No	8	11	No
160	16	28	No	138	143	No	8	10	No
500	14	29	No	136	150	No	11	9	No
1600	22	41	No	136	150	No	9	13	No
5000	21	27	No	10	171	Yes	10	9	No
Positive Control	104.2	1490	No	536	1592	No	316	153	No

Table 2a: Experiment 1 Plate incorporation assay Number of revertants per plate (mean of 3 plates) *solvent control with DMSO

Table 2b: Experiment 1 Plate incorporation assay Number of revertants per plate (mean of 3 plates)

	TA1537
Conc. (µg/plate)	— MA	+ MA	Cytotoxic (yes/no)
0*	5	13	No
50	0	11	No
160	2	15	No
500	2	10	No
1600	2	9	No
5000	0	3	Yes
Positive Control	84	109	No

*solvent control with DMSO

 

Table 3a: Experiment 2 Pre incubation assay Number of revertants per plate (mean of 3 plates)

	TA98			TA100			TA1535
Conc. (µg/plate)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)	— MA	+ MA	Cytotoxic (yes/no)
0*	19	25	No	121	116	No	12	9	No
250	21	25	No	97	102	No	7	11	No
500	20	30	No	98	91	No	13	11	No
1000	17	32	No	100	99	No	11	15	No
2000	18	31	No	101	110	No	8	10	No
4000	16	30	No	100	103	No	9	14	No
Positive Control	103	1628	No	608	1384	No	341	216	No

*solvent control with DMSO

Table 3b: Experiment 2 Pre incubation assay Number of revertants per plate (mean of 3 plates)

	TA1537
Conc. (µg/plate)	— MA	+ MA	Cytotoxic (yes/no)
0*	15	16	No
250	17	10	No
500	17	13	No
1000	17	15	No
2000	13	15	No
4000	12	13	No
Positive Control	263	131	No

*solvent control with DMSO

Conclusions:

On the basis of the above mentioned study ,The test substance did not show any reproducible mutagenic activity in any of the strains tested up to limit concentrations with and without metabolic activation in either the plate incorporation or the repeat preincubation assay. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

Executive summary:

In accordance with guideline OECD TG 471, a test of the genetic toxicity of 3-aminopropyl(triethoxy) (CAS number:919-30-2) was conducted using Salmonella typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 The test substance did not show any reproducible mutagenic activity in any of the strains tested up to limit concentrations with and without metabolic activation in either the plate incorporation or the repeat preincubation assay. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

Reference 4

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

data was taken from secondary source

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

(Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

Bacterial Reverse Mutation Assay was carried out for the RA chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Not specified

Target gene:

Histidine operon (S. typhimurium strains); tryptophan operon (E. coli).

Species / strain / cell type:

other: S. typhimurium, other: TA97, TA98 and TA100

Details on mammalian cell type (if applicable):

Not specified

Additional strain / cell type characteristics:

other: Not specified

Cytokinesis block (if used):

Not specified

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9

Test concentrations with justification for top dose:

8, 40, 200, 1000 and 5000 µg/plate

Vehicle / solvent:

Ethylene glycol dimethyl ether (EGDME)

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA100 without metabolic activation 100 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

TA1535 without metabolic activation 10000 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without metabolic activation 50 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA98, TA1538 without metabolic activation 1 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION

- Exposure duration: 2 days


NUMBER OF REPLICATIONS: five replicates

DETERMINATION OF CYTOTOXICITY
- Method: condition of background lawn

Rationale for test conditions:

Not specified

Evaluation criteria:

Not specified

Statistics:

Not specified

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations.

Vehicle controls validity:

valid

Untreated negative controls validity:

not valid

True negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No mutagenic potential was observed in any strain at any dose concentration

Remarks on result:

other: No mutagenic potential was observed in any strain at any dose concentration

Conclusions:

N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria in a study which was conducted according to OECD 471 but did not include TA102 or E.coli, compliant with GLP. No evidence of a test substance related increase in the number of revertants was observed with or without activation in the experiment. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

Executive summary:

Genetic toxicity study as per OECD guideline 471 (Bacterial Reverse Mutation Assay) was carried out for the RA chemical N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS number 1760-24-3).

During this test various strains were used TA98, TA100, TA1535, TA1537, and TA1538 at a dose range of 8, 40, 200, 1000 and 5000 µg/plate at metabolic activation factor Aroclor 1254 induced rat liver S9. Positive control substances are used methylmethanesulfonate,2-nitrofluorene,9-aminoacridine, and ethylmethanesulphonate by using the agar plate method.
After a complete analysis of the results the RA chemical which is structurally similar to test chemical
N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria in a study which was conducted according to OECD 471 but did not include TA102 or E.coli, compliant with GLP. No evidence of a test substance related increase in the number of revertants was observed with or without activation in the experiment. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

Reference 5

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Remarks:

Read across data

Adequacy of study:

weight of evidence

Study period:

Not Specified

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

secondary literature

Justification for type of information:

Data was taken from secondary source

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

bacterial reverse mutation assay

Principles of method if other than guideline:

Bacterial muation test was carried out for the test chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

Not Specified

Target gene:

Histidine operon (S. typhimurium strains); tryptophan operon (E. coli).

Species / strain / cell type:

other: TA98, TA100, TA1535, TA1537 and TA1538

Details on mammalian cell type (if applicable):

Not Specified

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

With metabolic activation: slight cytotoxicity in all strains at 2.5 and 5
mg/plate; Without metabolic activation: slight cytotoxicity in all strains at
2.5 and 5 mg/plate

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat liver S9

Test concentrations with justification for top dose:

0.1, 0.5, 1, 2.5 and 5 µg/plate

Vehicle / solvent:

DMSO

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

TA100 without metabolic activation 100 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

ethylmethanesulphonate

Remarks:

TA1535 without metabolic activation 10000 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

TA1537 without metabolic activation 50 µg/plate

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Remarks:

TA98, TA1538 without metabolic activation 1 µg/plate

Details on test system and experimental conditions:

Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation); preincubation

DURATION

- Exposure duration: 48 hours


NUMBER OF REPLICATIONS: triplicate plates, experiment repeated


DETERMINATION OF CYTOTOXICITY
- Method: other: condition of background lawn

Rationale for test conditions:

Not specified

Evaluation criteria:

Not specified

Statistics:

Not specified

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not specified

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

True negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

No mutagenic potential was observed in any strain at any dose concentration

Remarks on result:

other: No mutagenic potential was observed in any strain at any dose concentration

Conclusions:

N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria, in a study which was conducted according to OECD 471, compliant with GLP. No evidence of a test substance related increase in the number of revertants was observed with or without metabolic activation in the initial or repeat experiment. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

Executive summary:

Genetic toxicity study as per OECD guideline 471 (Bacterial Reverse Mutation Assay) was carried out for the RA chemical N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS number 1760-24-3).

During this test various strains were used TA98, TA100, TA1535, TA1537, and TA1538 at a dose range of 0.1, 0.5, 1, 2.5, and 5 µg/plate at metabolic activation factor Aroclor 1254 induced rat liver S9. Positive control substances are used methylmethanesulfonate,2-nitrofluorene,9-aminoacridine, and ethylmethanesulphonate by using the agar plate method.
After a complete analysis of the results the RA chemical which is structurally similar to test chemical N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria, in a study which was conducted according to OECD 471, compliant with GLP. No evidence of a test substance-related increase in the number of revertants was observed with or without metabolic activation in the initial or repeat experiment. It is concluded that the substance is negative for mutagenicity to bacteria under the conditions of the test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Ames assay:

Study 1: Genetic toxicity study as per OECD guideline 471 (Bacterial Reverse Mutation Assay) was carried out for the RA chemical N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS number 1760-24-3). During this test various strains were used TA98, TA100, TA1535, TA1537, and TA1538 at a dose range of 0.1, 0.5, 1, 2.5, and 5 µg/plate at metabolic activation factor Aroclor 1254 induced rat liver S9. Positive control substances are used methylmethanesulfonate,2-nitrofluorene,9-aminoacridine, and ethylmethanesulphonate by using the agar plate method. After a complete analysis of the results the RA chemical which is structurally similar to test chemical N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria, in a study which was conducted according to OECD 471, compliant with GLP. No evidence of a test substance-related increase in the number of revertants was observed with or without metabolic activation in the initial or repeat experiment. It is concluded that the substance is negative for mutagenicity to bacteria under the conditions of the test.

 

Study 2: Genetic toxicity study as per OECD guideline 471 (Bacterial Reverse Mutation Assay) was carried out for the RA chemical N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS number 1760-24-3). During this test various strains were used TA98, TA100, TA1535, TA1537, and TA1538 at a dose range of 8, 40, 200, 1000 and 5000 µg/plate at metabolic activation factor Aroclor 1254 induced rat liver S9. Positive control substances are used methylmethanesulfonate,2-nitrofluorene,9-aminoacridine, and ethylmethanesulphonate by using the agar plate method. After a complete analysis of the results the RA chemical which is structurally similar to test chemical N-(3-(trimethoxysilyl)propyl)ethylenediamine has been tested for mutagenicity to bacteria in a study which was conducted according to OECD 471 but did not include TA102 or E.coli, compliant with GLP. No evidence of a test substance related increase in the number of revertants was observed with or without activation in the experiment. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

 

Study 3: In accordance with guideline OECD TG 471, a test of the genetic toxicity of 3-aminopropyl(triethoxy) (CAS number:919-30-2) was conducted using Salmonella typhimurium strains TA 98, TA 100, TA 1535, and TA 1537 The test substance did not show any reproducible mutagenic activity in any of the strains tested up to limit concentrations with and without metabolic activation in either the plate incorporation or the repeat preincubation assay. It is concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.

 

In vitro mammalian chromosome aberration study:

In vitro mammalian chromosomal aberration test was performed to determine the mutagenic nature of the test chemical. The test chemical was mixed with DMSO and used at dose level of 0, 0.000001, 0.00001, 0.000, or 0.001 M using pseudo-diploid Chinese hamster cell line (Don). Three hours after 1.0-1.2 X 106 cells per TD-40 culture bottle were seeded, BUdR (1µg/ml) and test chemical was added to the cultures under an ordinary yellow darkroom safety lamp. Concurrent solvent control was also included in the study. All cultures were kept in complete darkness at 37° C for 26 hours (this covered two rounds of cell cycle), and 0.25µg colchicine/ml was added for the final 2 hours. The cells were collected by scraping them with a rubber policeman and prepared air-dried slides following hypotonic treatment and fixation in ice-cold methanol: acetic acid (3: 1). The chromosome slide was stained in aqueous solution of 33258 Hoechst for 10 minutes, rinsed briefly in tap water, and mounted in phosphate buffer (pH 7.0) with a cover slip. The slide was exposed to an electric light (60 W, at 12-cm distance) for 1 hour. The cover slip was removed by tap water, and the slide was incubated in 1 M NaH2P04 (pH 8.0, 83-85° C) for 10 minutes, rinsed, and stained in 2.5% Giemsa (in phosphate buffer, 0.07 M, pH 7.0) for 5 minutes. Conventional Giemsa-stained slides were also prepared for scanning of chromosome aberrations. The frequency of aberrations, excluding gaps, was indicated by the number of breaks per cell. A ring, a dicentric, and a chromatid exchange were each scored as two breaks, a tricentric as four breaks, and an acentric fragment or an isochromatid break as one break. The test chemical did not induce chromosome aberration in pseudo-diploid Chinese hamster cell line (Don) and hence it is not likely to classify as a gene mutant in vitro.

 

In vitro gene mutation study in mammalian cells:

The test chemical was assayed for gene mutations in the Chinese Hamster Ovary (CHO) cells both in the absence and presence of S9 metabolic activation. Positive (dimethylnitrosamine with S9 activation and ethylmethanesulfonate without S9 activation) and vehicle control (cell culture medium and DMSO) materials were tested concurrently to assure both the sensitivity of the test system. The study was performed according to OECD 476 Guidelines. The test concentrations used were- 0.1 to 4.0 mg/ml without S9 metabolic activation and 2.0 to 5.0 mg/ml with S9 activation. did not product any statistically significant increases in the incidence of mutations of CHO cells within a range of cytotoxic-to-noncytotoxic concentrations between 2.5 to 4.0 mg/ml in tests without an S9 metabolic activation system. With S9 activation, one intermediate dose of 2.5 mg/ml produced a mutant incidence in one of the two dosed cultures which was statistically greater than the concurrent controls. No dose-related trend in mutant values was observed in the test with or without S9 activation. The biological significance of the single increase was evaluated by determining reproducibility in an independent repeat test over a narrower range of concentrations with S9 activation. No significant or dose-related increases were observed in the repeat test. The test substance did not induce any statistical significant increase in the number of cells in the presence or absence of the S9 metabolic activation. Hence, the test substance can be considered to be non-mutagenic.

Justification for classification or non-classification

Based on all the available RA data it can be concluded that the given test chemical has structural similarity with the RA substance. It can be concluded that the test substance is negative for mutagenicity to bacteria under the conditions of the test.